舒尼替尼经线粒体内活性氧引起心肌细胞凋亡

Mitochondrial reactive oxygen species in sunitinib induced cardiomyocytes apoptosis

目的探讨舒尼替尼引起心肌细胞凋亡的作用机制。方法 H9C2细胞分别用舒尼替尼0.1,1和10μmol.L-1处理24,48,72 h,MTT法测定细胞存活率;流式细胞仪分别测定处理24 h细胞的凋亡、细胞内活性氧(ROS)、线粒体膜电位(ΔΨm)水平及胱天蛋白酶3活性。结果与同一时间点正常对照组相比,舒尼替尼1,10μmol.L-1处理24,48,72 h后,细胞存活率分别明显下降了22%和32%(24 h);41%和68%(48 h);62%和82%(72 h)(P<0.05)。与正常对照组相比,舒尼替尼1,10μmol.L-1作用24 h后,心肌细胞内ROS水平显著升高(4.41±0.76 vs 8.68±0.74,3.57±1.45)(P<0.05),线粒体膜电位下降(309±6 vs 244±28,174±2)(P<0.05),胱天蛋白酶3活性升高(0.96±0.13 vs 59.40±13.17,79.90±0.06)(P<0.05)及细胞凋亡率增加(〔6.03±0.40)%vs(21.05±5.55)%,(59.05±4.62)%〕(P<0.05)。结论舒尼替尼可通过诱导心肌细胞内ROS的产生,经线粒体内途径引起心肌细胞凋亡。

OBJECTIVE To investigate the cardiac toxicity of sunitinib.METHODSH9C2 cells were exposed to sunitinib 0.1,1 and 10 μmol·L^-1 for 24,48 and 72 h.Cell viability was determined by MTT assay.Apoptosis,the level of intracellular reactive oxygen species（ROS）,mitochondrial membrane potential（ΔΨm） and caspase 3 activity were detected by flow cytometry in the 24 h-treatment group.RESULTS Compared with normal control group,sunitinib 1 and 10 μmol·L^-1 significantly decreased cells viability by 22% and 32% for 24 h-treatment group,by 48% and 68% for 48-h treatment group,and by 62% and 82% for 72 h-treatment group（P0.05）.Compared with normal control group,sunitinib 1 and 10 μmol·L^-1 significanty increased the level of intracellular ROS（4.41±0.76 vs 8.68±0.74,3.57±1.45;P0.05）,significantly decreased the ΔΨm（309±6 vs 244±28,174±2）（P0.05）,increased caspase 3 activity（0.96±0.13 vs 59.40±13.17,79.90±0.06）（P0.05）,and decreased cell apoptosis（（6.03±0.40）% vs（21.05±5.55）%,（59.05±4.62）%）（P0.05）.CONCLUSIONSunitinib could prompt the apoptosis of H9C2 cells through mitochondrial pathway by generating intracellular ROS.